Protein Kinase C Protocols

Protein Kinase C Protocols

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Since the discovery that protein kinase C (PKC) transduces the ab- dance of signals that result in phospholipid hydrolysis, this enzyme has been at the forefront of research in signal transduction. Protein Kinase C Protocols covers fundamental methods for studying the structure, function, regulation, subcellular localization, and macromolecular interactions of PKC. Protein Kinase C Protocols is divided into 11 sections representing the major aspects of PKC regulation and function. Part I contains an introduction and a historical perspective on the discovery of PKC by Drs. Yasutomi Nishizuka and Ushio Kikkawa. Part II describes methods to purify PKC. Part III describes the standard methods for measuring PKC activity: its enzymatic activity and its stimulus-dependent translocation from the cytosol to the membrane. Part IV describes methods for measuring the membrane interaction of PKC in vivo and in vitro. Part V provides methodologies and techniques for measuring the ph- phorylation state of PKC, including a protocol for measuring the activity of PKC's upstream kinase, PDK-1. Novel methods for identifying substrates are described in Part VI. Part VII presents protocols for expressing and analyzing the membrane targeting domains of PKC. Part VIII provides a comprehensive c- pilation of methods used to identify binding partners for PKC. Part IX describes pharmacological probes used to study PKC. The book ends with a presentation of genetic approaches to study PKC (Part X) and a discussion of approaches used to study PKC in disease (Part XI).show more

Product details

  • Hardback | 584 pages
  • 158 x 232 x 32mm | 920.81g
  • Humana Press Inc.
  • Totowa, NJ, United States
  • English
  • 2003 ed.
  • biography
  • 1588290689
  • 9781588290687
  • 2,394,838

Review quote

"...a comprehensive collection of established and emerging techniques used for the study of protein kinase C, and is quite simply an essential guide for any lab interested in cell impressive collection of expert contributors...this is an excellent lab manual which provides very detailed and easy to follow methods and instructions that would be useful for any lab that is interested in any form of cell signalling..." - Proteomics "...a valuable book for everybody interested in PKC, and it is a must for everybody doing research with it." - ChemBioChemshow more

Back cover copy

Protein kinase C impacts on nearly every aspect of signal transduction, placing it at the forefront of research in how cells transduce information. In Protein Kinase C Protocols, leading investigators from academia and industry present a comprehensive collection of classical and emerging laboratory techniques for the study of this influential protein, all optimized for successful and reproducible results. Described in step-by-step detail, these methods can be easily used to explore the structure, function, regulation, subcellular localization, and macromolecular interactions of protein kinase C. Included in this collection are easy-to-follow techniques to purify protein kinase C, to measure its activity and its membrane interaction in vivo and in vitro, and to identify substrates. There are also methodologies and techniques for measuring the phosphorylation state of protein kinase C, for expressing and analyzing the membrane-targeting domains of protein kinase C, and for detecting binding proteins. Additional chapters address pharmacological probes, genetic approaches, and protein kinase C in disease. Each protocol is introduced in the context of protein kinase C function and regulation and contains many notes on how best to deal with the problems that may occur. Comprehensive and authoritative, Protein Kinase C Protocols offers a timely compilation of biophysical, biochemical, cell biological, and molecular biological approaches that will ensure successful results in every laboratory working in this rapidly advancing more

Table of contents

Part I. Introduction The Ins and Outs of Protein Kinase C Alexandra C. Newton Early Studies of Protein Kinase C: A Historical Perspective Yasutomi Nishizuka and Ushio Kikkawa Part II. Expression and Purification Expression and Purification of Protein Kinase C from Insect Cells Hideyuki Mukai and Yoshitaka Ono Expression and Purification of Protein Kinase Cd from Bacteria Michael Gschwendt and Luise Stempka Part III. Measuring the Activity of Protein Kinase C Complexities in Protein Kinase C Activity Assays: An Introduction Julianne J. Sando Enzyme Assays for Protein Kinase C Activity Julianne J. Sando and Jacquelyn K. Beals Subcellular Translocation of Protein Kinase C Julianne J. Sando, Jacquelyn K. Beals, and Isa M. Hussaini Part IV. Measuring the Interaction of Protein Kinase C with Membranes Measuring the Interaction of Protein Kinase C with Membranes: An Introduction Alexandra C. Newton Fluorescence Imaging of Protein Kinase C Translocation in Living Cells Naoaki Saito Measuring the Binding of Protein Kinase C to Sucrose-Loaded Vesicles Jennifer Giorgione and Alexandra C. Newton Use of Stopped-Flow Fluorescence Spectroscopy to Measure Rapid Membrane Binding by Protein Kinase C Eric A. Nalefski and Alexandra C. Newton [3H]Phorbol 12,13-Dibutyrate Binding Assay for Protein Kinase C and Related Proteins Nancy E. Lewin and Peter M. Blumberg Part V. Measuring the Phosphorylation of Protein Kinase C Protein Kinase C Phosphorylation: An Introduction Peter J. Parker Pulse-Chase Analysis of Protein Kinase C Mikiko Takahashi and Yoshitaka Ono PDK-1 and Protein Kinase C Phosphorylation Alex Toker In Vitro Autophosphorylation of Protein Kinase C Isozymes Antonio M. Pepio and Wayne S. Sossin Tyrosine Phosphorylation of Protein Kinase C Toshiyoshi Yamamoto, Emiko Yamauchi, Hisaaki Taniguchi, Hidenori Matsuzaki, and Ushio Kikkawa Methods to Study Dephosphorylation of Protein Kinase C In Vivo Karen England and Martin Rumsby Phosphopeptide-Specific Antibodies to Protein Kinase C Wayne S. Sossin Part VI. Identifying Protein Kinase C Substrates Identifying Protein Kinase C Substrates: An Introduction Alex Toker A Chemical Genetic Approach for the Identification of Direct Substrates of Protein Kinases Kavita Shah and Kevan M. Shokat Studying the Optimal Peptide Substrate Motifs of Protein Kinase C Using Oriented Peptide Libraries Michael B. Yaffe Part VII. Structural Analysis of Protein Kinase C Structural Analysis of Protein Kinase C: An Introduction James H. Hurley Bacterial Expression and Purification of C1 and C2 Domains of Protein Kinase C Isoforms Wonhwa Cho, Michelle Digman, Bharath Ananthanarayanan, and Robert V. Stahelin Crystallization of the Protein Kinase Cd C1B Domain Gongyi Zhang and James H. Hurley Part VIII. Methods for Detecting Binding Proteins Methods for Detecting Binding Proteins: An Introduction Gerda Endemann and Daria Mochly-Rosen Detection of Protein Kinase-Binding Partners by the Yeast Two-Hybrid Analysis Betty Y. Chang and Christine A. Cartwright Glutathione S-Transferase Pull-Down Assay Deborah Schechtman, Daria Mochly-Rosen, and Dorit Ron Overlay Method for Detecting Protein-Protein Interactions Deborah Schechtman, Christopher Murriel, Rachel Bright, and Daria Mochly-Rosen An Overlay Assay for Detecting Protein Kinase C-Binding Proteins and Substrates Susan Jaken Functional Proteomic Analysis of the Protein Kinase C Signaling System Jason M. Pass, Junshow more

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